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IRG1 Recombinant Rabbit Monoclonal Antibody [PSH07-31]

Usd: 385

Specification

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Catalog# HA722819

IRG1 Recombinant Rabbit Monoclonal Antibody [PSH07-31]

Application

  • WB

  • IF-Cell

Reactivity

  • Mouse

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

IRG1 Recombinant Rabbit Monoclonal Antibody [PSH07-31]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within mouse IRG1 aa 1-488.

Species Reactivity

Mouse

Validated Applications

WB, IF-Cell

Molecular Weight

Predicted band size: 54 kDa

Positive Control

RAW264.7 treated with 0.1μg/mL LPS for 6 hours cell lysate, J774A.1 treated with 1μg/mL LPS for 24 hours cell lysate, mBMMC treated with 0.1μg/mL LPS and 10ng/mL IFN-gamma for 24 hours cell lysate, mBMSC treated with 40ng/mL MCSF for 7 days then add 100ng/mL LPS for 48 hours cell lysate, RAW264.7 cells treated with 0.1μg/mL LPS for 6 hours.

Conjugation

unconjugated

Clone Number

PSH07-31

Reactivity Data

WB IF-Cell
Mouse
Tested Tested
Tested Tested
Human
Not Recommended
Not Recommended
Rat
Not Recommended
Not Recommended

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

Target

Function

Cis-aconitate decarboxylase that catalyzes production of itaconate and is involved in the inhibition of the inflammatory response. Acts as a negative regulator of the Toll-like receptors (TLRs)-mediated inflammatory innate response by stimulating the tumor necrosis factor alpha-induced protein TNFAIP3 expression via reactive oxygen species (ROS) in LPS-tolerized macrophages. Involved in antimicrobial response of innate immune cells; ACOD1-mediated itaconic acid production contributes to the antimicrobial activity of macrophages by generating itaconate, leading to alkylation of proteins, such as TFEB. Involved in antiviral response following infection by flavivirus in neurons: ACOD1-mediated itaconate production inhibits the activity of succinate dehydrogenase, generating a metabolic state in neurons that suppresses replication of viral genomes (By similarity). Plays a role in the embryo implantation (By similarity).

Background References

1. Wu R et al. The Dual Role of ACOD1 in Inflammation. J Immunol. 2023 Aug

2. Wu R et al. Mitochondrial ACOD1/IRG1 in infection and sterile inflammation. J Intensive Med. 2022 Feb

Subcellular Location

Mitochondrion.

UNIPROT

SwissProt: P54987 Mouse

Synonyms

ACOD1 antibody

Aconitate decarboxylase 1 antibody

Aconitate decarboxylase antibody

CAD antibody

Cis aconitate decarboxylase antibody

Immune responsive gene 1 protein homolog antibody

Immune-responsive gene 1 protein homolog antibody

Immunoresponsive 1 homolog (mouse) antibody

Irg1 antibody

IRG1_HUMAN antibody

Expand

Images

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Western blot analysis of IRG1 on different lysates with Rabbit anti-IRG1 antibody (<a href="/products/HA722819" style="font-weight: bold;text-decoration: underline;">HA722819</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.<br /><br />Lane 1: RAW264.7 cell lysate<br />Lane 2: RAW264.7 treated with 0.1μg/mL LPS for 6 hours cell lysate<br />Lane 3: J774A.1 cell lysate<br />Lane 4: J774A.1 treated with 1μg/mL LPS for 24 hours cell lysate<br />Lane 5: mBMMC cell lysate<br />Lane 6: mBMMC treated with 0.1μg/mL LPS and 10ng/mL IFN-gamma for 24 hours cell lysate<br />Lane 7: mBMSC cell lysate<br />Lane 8: mBMSC treated with 40ng/mL MCSF for 7 days then add 100ng/mL LPS for 48 hours cell lysate<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 54 kDa<br />Observed band size: 54 kDa<br /><br />Exposure time: 10 seconds; ECL: K1801;<br /><br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/HA722819" style="font-weight: bold;text-decoration: underline;">HA722819</a>) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    ☑ Cell treatment (CT)

    Western blot analysis of IRG1 on different lysates with Rabbit anti-IRG1 antibody (HA722819) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution.

    Lane 1: RAW264.7 cell lysate
    Lane 2: RAW264.7 treated with 0.1μg/mL LPS for 6 hours cell lysate
    Lane 3: J774A.1 cell lysate
    Lane 4: J774A.1 treated with 1μg/mL LPS for 24 hours cell lysate
    Lane 5: mBMMC cell lysate
    Lane 6: mBMMC treated with 0.1μg/mL LPS and 10ng/mL IFN-gamma for 24 hours cell lysate
    Lane 7: mBMSC cell lysate
    Lane 8: mBMSC treated with 40ng/mL MCSF for 7 days then add 100ng/mL LPS for 48 hours cell lysate

    Lysates/proteins at 10 µg/Lane.

    Predicted band size: 54 kDa
    Observed band size: 54 kDa

    Exposure time: 10 seconds; ECL: K1801;

    4-20% SDS-PAGE gel.

    Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722819) at 1/1,000 dilution and competitor's antibody at 1/1,000 dilution were used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Immunocytochemistry analysis of RAW264.7 cells treated with 0.1μg/mL LPS for 6 hours labeling IRG1 with Rabbit anti-IRG1 antibody (<a href="/products/HA722819" style="font-weight: bold;text-decoration: underline;">HA722819</a>) at 1/100 dilution.<br /><br />Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IRG1 antibody (<a href="/products/HA722819" style="font-weight: bold;text-decoration: underline;">HA722819</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    ☑ Cell treatment (CT)

    Immunocytochemistry analysis of RAW264.7 cells treated with 0.1μg/mL LPS for 6 hours labeling IRG1 with Rabbit anti-IRG1 antibody (HA722819) at 1/100 dilution.

    Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-IRG1 antibody (HA722819) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

    Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"